Platelet-reactive sites in human collagens I and III: evidence for cell-recognition sites in collagen unrelated to RGD and like sequences.

Following fragmentation of the collagen molecule with cyanogen bromide (CB), two major platelet-aggregatory sites were detected with human platelets in the alpha 1(I)-chain of human collagen I corresponding to those detected previously in bovine alpha 1(I)-chains. Two main sites were also detected in the human alpha 1(III)-chain, at locations different from those in the alpha 1(I)-chain. Only one of these had been previously recognised. The new site was found in the peptide alpha 1(III)CB3, the amino acid sequence of which does not contain the cell-recognition site RGD nor comparable sequences that might be supposed to serve this function such as KGD, RGE or KGE. The peptide does, however, contain the sequence GRPGRPGER which reflects a spacing of basic residues (at positions 2 and 9) we have previously postulated to be essential for collagen to cause platelet aggregation. None of the CB-derived peptides was able to cause an aggregation of rabbit platelets. Human platelet secretion, as aggregation, was only induced by CB-derived fragments in triple-helical, polymeric form. One fragment, peptide alpha 1(III)CB8, was able to induce secretion although lacking aggregatory activity. Platelet adhesion occurred to all of the fragments, including those lacking aggregatory activity. Adhesion also occurred to the collagen-like polypeptide (PGP) n. However, inhibition studies suggested that the GPP sequence which occurs frequently along the length of the collagen molecule is not responsible for platelet adhesion to collagen.